Simultaneous surveillance of 30 phytopathogens in potato using a high-throughput diagnostic system combining...

Global potato production suffers severe yield losses owing to phylogenetically diverse pathogens spanning fungi, bacteria, viruses, viroids, nematodes and insects. These biotic stressors cause significant annual yield losses in major potato-growing regions, posing an urgent threat to global food security. Current molecular diagnostics remain constrained by limited multiplex capacity (typically <10 targets/assay) and protracted workflows, fundamentally impeding comprehensive phytosanitary management.

Our integrated diagnostic platform synergizes multiplex PCR and nucleic acid-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to achieve unprecedented 30-plex detection of 30 high-risk potato pathogens including six fungi, four bacteria, 16 viruses, two viroids, one nematode and one insect within 10 h. The platform demonstrates 6-60 copy sensitivity across all targets via optimized dual-reaction primer pooling, enabling pathogen identification in extracted DNA/RNA from symptomatic potato tissues using SpectroCHIP arrays with 96-sample format. Crucially, this platform eliminates cross-reactivity through mass-resolved resolution, successfully detected pathogens in potato tissues, overcoming the limitation of conventional current PCR assays.

This diagnostic breakthrough achieves 10-fold higher multiplexing capacity than conventional PCR assays and cost reduction through multiplex optimization, enabling simultaneous detection of 30 pathogens per assay. High-throughput sample processing can be performed with 96- or 384-well SpectroCHIP arrays, demonstrating scalable screening capacity for precision agriculture. The platform's cost-effective consumables and rapid workflow enable instant phytosanitary monitoring in seed certification programs and transnational quarantine protocols. © 2025 Society of Chemical Industry.